Host-specific recognition kairomone for the parasitoidMicroplitis croceipes (Cresson)

A water-extractable host recognition kairomone in frass of corn earworm,Helicoverpa zea Boddie (Lepidoptera: Noctuidae), host larvae stimulates antennation by females of the parasitoidMicroplitis croceipes Cresson (Braconidae: Hymenoptera). In addition, when the wasps contact water extracts of host frass they will subsequently fly in a flight tunnel to odor associated with the extract. Contact with water extracts of cowpea leaves or with water extracts of frass from larvae of nonhost beet armyworm, fall armyworm, or cabbage looper that were fed cowpea leaves does not stimulate antennation, nor do wasps fly to associated odors after contact with these substances. However, contact with the water extract of host frass in association with hexane extract of cowpea-fed nonhost frass will induce the wasps to subsequently fly to the hexane extract of the nonhost frass when it is used as an odor source in a flight tunnel. Thus the host-specific kairomone by whichM. croceipes recognizes the frass of its host is extractable with water. This substance plays a crucial role in the foraging behavior of this parasitoid by allowing it to recognize host frass and to learn to search for odors originating from plants on which the host is feeding.     

Identification and Synthesis of a Kairomone Mediating Host Location by Two Parasitoid Species of the Cassava Mealybug Phenacoccus herreni

Two encyrtid species, Acerophagus coccois and Aenasius vexans, parasitoids of the cassava mealybug Phenacoccus herreni use a contact kairomone from the body surface of their host as a host-location stimulant. The kairomone was synthesized and identified as O-caffeoylserine based on a combination of chromatographic methods. The synthetic compound was determined to be active.     

Identification and synthesis of a kairomone inducing oviposition by parasitoidAphytis melinus from California red scale covers

The parasitoid waspAphytis melinus uses a kairomone from the cover of its scale host, California red scale (Aonidiella aurantii), as an oviposition stimulant. The kairomone was isolated from extracts of scale covers, and identified asO-caffeoyltyrosine by a combination of spectroscopic methods. The kairomone was synthesized, and the synthetic compound was determined to be as active as the chemical isolated from scale covers.     

Antennal electrophysiological responses of three parasitic wasps to caterpillar-induced volatiles from maize (Zea mays mays), cotton (Gossypium herbaceum), and cowpea (Vigna unguiculata)

Many parasitic wasps are attracted to volatiles that are released by plants when attacked by potential hosts. The attractiveness of these semiochemicals from damaged plants has been demonstrated in many tritrophic systems, but the physiological mechanisms underlying the insect responses are poorly understood. We recorded the antennal perception by three parasitoids (Cotesia marginiventris, Microplitis rufiventris, and Campoletis sonorensis) to volatiles emitted by maize, cowpea, and cotton plants after attack by the common caterpillar pest Spodoptera littoralis. Gas chromatography-electroantennography (GC-EAG) recordings showed that wasps responded to many, but not all, of the compounds present at the physiologically relevant levels tested. Interestingly, some minor compounds, still unidentified, elicited strong responses from the wasps. These results indicate that wasps are able to detect many odorant compounds released by the plants. It remains to be determined how this information is processed and leads to the specific behavior of the parasitoids.     

Aggregation pheromone ofCarpophilus antiquus (Coleoptera: Nitidulidae) and kairomonal use ofC. lugubris pheromone byC. antiquus

Males ofCarpophilus antiquus Melsheimer (Coleoptera: Nitidulidae) emit an aggregation pheromone that was found to be a novel hydrocarbon, (3E,5E,7E,9E)-6,8-diethyl-4-methyl-3,5,7,9-dodecatetraene. A synthetic scheme and spectra (mass and proton NMR) are given for the compound. Beetles produced the pheromone when feeding on a variety of media, including the brewer's yeast-based artificial diet, fermenting whole-wheat bread dough, corn, and prunes; live baker's yeast was generally added to the food media. Males held individually produced, on average, 25 × more pheromone per beetle than males held in groups of 10 or more. Pheromone was not produced until males were at least 5 days old but was still detected from the oldest beetles tested (47 days). In field tests, the pheromone was attractive to both sexes ofC. antiquus, and it was synergized by food volatiles: A combination of pheromone and fermenting whole wheat dough attracted 2.5× more beetles than pheromone alone, but dough by itself was not significantly more attractive than the control. Semiochemical interactions were studied amongC. antiquus and two other sympatric species for which pheromones are known,C. lugubris Murray andC. freemani Dobson.C. antiquus responded readily to the pheromone ofC. lugubris, but all other interspecific responses to the pheromones were weak. In a sample of naturally infested corn ears, the presence ofC. antiquus was strongly associated with the presence ofC. lugubris, as would be expected if the pheromone ofC. lugubris serves as a kairomone forC. antiquus.     

Mite predator responses to prey and predator-emitted stimuli

We found that the searching behavior of two acarine predators,Amblyseius fallacis andPhytoseiulus macropilis, for prey,Tetranychus urticae, is affected by the following stimuli: (1) prey silk and associated feces, whose combined physical and chemical properties elicit reduction in the rate of predator movements and longer halts; (2) kairomone extracted from prey silk and associated feces, which, upon contact, elicits frequent predator return to prey-inhabited locales; and (3) predator-emitted marking pheromone, which elicits shorter duration of search in presearched prey locales. We also found that treatment of filter paper with prey kairomone or silk enhanced predator location of prey eggs, leading us to speculate that application of synthetic prey kairomone could be useful in pest management programs.     

Interspecific response to sex pheromones,and calling behavior of severalAttagenus species (Coleoptera: Dermestidae)

Females of severalAttagenus species demonstrated calling behaviors similar to those previously reported forA. elongatulus. Attagenus rufipennis females did not call until they were 12–14 days old. OtherAttagenus species called at approx. 4 days of age. All species exhibited diurnal periods of calling activity. Interspecific pheromone responses were tested, withA. megatoma megatoma, A. megatoma canadensis, A. megatoma japonicus, andA. schaefferi spurcus showing equal cross-responses. Male response to females was demonstrated inA. rufipennis andA. bicolor. Attagenus rufipennis, bicolor, andelongatulus males responded only to female extracts of their own species.Mention of a proprietary product does not constitute an endorsement by the USDA.     

Responses of parasiteEdovum puttleri to kairomone from eggs of colorado potato beetle,Leptinotarsa decemlineata

When females of the eulophid parasiteEdovum puttleri were exposed to ann-hexane wash of eggs of the Colorado potato beetle,Leptinotarsa decemlineata, their responses indicated that the eggs contained a kairomone which elicited host-finding and host-acceptance behaviors. The hostfinding responses included an increased amount of time searching glass beads treated with kairomone, and drumming on the beads with the flagellum of the antennae. Host-acceptance responses included grasping of the beads with the legs, raising of the abdomen, and probing the kairomone-treated surface with the ovipositor. None of these responses were noted in females on beads treated only with hexane, or in males exposed to kairomone-treated beads. When kairomone was applied to a flat surface, filter paper disks, few episodes of drumming and no episodes of probing with the ovipositor were noted. Responses ofE.puttleri to eggs of Colorado potato beetle were similar to those on kairomone-treated beads, but females normally drilled into eggs and fed on host fluids after retracting the ovipositor. The kairomone is volatile, and responsiveness was reduced in parasites exposed to beads treated several hours previously.Hymenoptera: Eulophidae.Coleoptera: Chrysomelidae.     

Beneficial arthropod behavior mediated by airborne semiochemicals. II. Olfactometric studies of host location by the parasitoidMicroplitis croceipes (Cresson) (Hymenoptera: Braconidae)

The response ofMicroplitis croceipes (Cresson) to odors from a plant-host complex was investigated using a four-choice olfactometer. Female, but not male, parasitoids responded in a dose-dependent fashion to volatiles from the plant-host complex and oviposition experience enhanced this response. Female age had no apparent effect on the response. Both artificially damaged leaves and frass elicited positive responses but of lower magnitude than those elicited by the plant-host complex. Volatiles collected from the plant-host complex placed on filter paper also elicited positive responses by female parasitoids.Mention of a commercial or proprietary product does not constitute an endorsement by the USDA.     

Pheromone chirality of african palm weevil,Rhynchophorus phoenicis (F.) and palmetto weevil,Rhynchophorus cruentatus (F.) (Coleoptera: Curculionidae)

There are four stereoisomers of both 3-methyl-octan-4-ol, the aggregation pheromone of the African palm weevil,Rhynchophorus phoenicis (F.) and 5-methyl-octan-4-ol, the aggregation pheromone of the palmetto weevil,Rhynchophorus cruentatus (F.). Synthetic stereoisomers of 3-methyl-octan-4-ol and 5-methyl-octan-4-ol were baseline-separated on a Cyclodex-B fused silica column. Use of this column in gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric (GC-MS) analyses revealed that only one stereoisomer, (3S,4S)-3-methyl-octan-4-ol and (4S,5S)-5-methyl-octan-4-ol, is produced by maleR. phoenicis and maleR. cruentatus, respectively, and elicits good antennal responses by conspecific male and female weevils. In field trapping experiments, withR. phoenicis in Côte d'Ivoire andR. cruentatus in Florida, (3S,4S)-3-methyl-octan-4-ol and (4S,5S)-5-methyl-octan-4-ol strongly enhanced attraction of fresh palm tissue, whereas other stereoisomers were behaviorally benign. Stereoisomeric 3-methyl-octan-4-ol and 5-methyl-octan-4-ol may be utilized to monitor and/or manage populations of these two palm weevils.     

Plant–herbivore–carnivore Interactions in Cotton, <Emphasis Type="Italic">Gossypium hirsutum</Emphasis>: Linking Belowground and Aboveground

Most studies on plant–herbivore interactions focus on either root or shoot herbivory in isolation, but above- and belowground herbivores may interact on a shared host plant. Cotton (Gossypium spp.) produces gossypol and a variety of other gossypol-like terpenoids that exhibit toxicity to a wide range of herbivores and pathogens. Cotton plants also can emit herbivore-induced volatile compounds at the site of damage and systemically on all tissues above the site of damage. As these volatile compounds attract natural enemy species of the herbivore, they are thought to represent an indirect plant defense. Our study quantified gossypol and gossypol-like compounds in cotton plants with foliage feeding (Heliocoverpa zea), root feeding (Meloidogyne incognita), or their combination. Cotton plants with these treatments were studied also with respect to induced local and systemic volatile production and the attraction of the parasitic wasp Microplitis croceipes to those plants. We also evaluated whether foliage or root feeding affected foliar nitrogen levels in cotton. After 48 hr of leaf feeding and 5 wk of root feeding, local and systemic induction of volatiles (known to attract parasitoids such as M. croceipes) occurred with herbivore damage to leaves, and it increased in levels when root herbivory was added. Nevertheless, M. croceipes were equally attracted to plants with both leaf and root damage and leaf damage only. In contrast to previous studies in cotton, production of gossypol and gossypol-like compounds was not induced in leaf and root tissue following foliage or root herbivory, or their combination. We conclude that root feeding by M. incognita has little influence on direct and indirect defenses of Gossypium hirsutum against insect herbivory.     

Oviposition stimulants inBarbarea vulgaris forPieris rapae andP. napi oleracea: isolation,identification and differential activity

The closely related butterflies,Pieris rapae andP. napi oleracea, readily laid eggs onBarbarea vulgaris in greenhouse cages. When offered a choice between cabbage andB. vulgaris, P. rapae showed no preference, butP. napi oleracea preferredB. vulgaris. Bioassays of extracts ofB. vulgaris foliage revealed the presence of oviposition deterrent(s) in l-butanol extracts as well as stimulants in the postbutanol water extracts. However, the deterrent effect was apparently outweighed by the strong stimulatory effect in the whole plants. The postbutanol water extract was preferred over an equivalent cabbage extract by both species, but more significantly in the case ofP. napi oleracea. The stimulants were isolated by open column chromatography and HPLC, and the activity was associated with three glucosinolates.P. napi oleracea was more sensitive thanP. rapae to the natural concentration of compounds1 and3, whereas both species were strongly stimulated to oviposit by natural concentrations of compound2. Compounds1 and2 were identified as (2R)-glucobarbarin and (2S)-glucobarbarin, respectively, and3 was identified as glucobrassicin, on the basis of their UV, mass, and NMR spectra. When the pure compounds were tested at the same concentrations applied to bean plants, the (2R)-glucobarbarin at 0.2 mg/plant was preferred over a standard cabbage extract by both butterfly species. However, at a dose of 0.02 mg/plant,P. rapae preferred the cabbage extract whereasP. napi oleracea still preferred the (2R)-glucobarbarin. No such difference in response of the two species to the same two concentrations of (2S)-glucobarbarin was obtained. The results indicate a distinct difference in sensitivity of these butterflies to the epimers of glucobarbarin, and the differences in behavioral responses of the two butterfly species depend to a large extent on the concentration of stimulant present.     

The Role of Fresh <Emphasis Type="Italic">versus</Emphasis> Old Leaf Damage in the Attraction of Parasitic Wasps to Herbivore-Induced Maize Volatiles

The odor produced by a plant under herbivore attack is often used by parasitic wasps to locate hosts. Any type of surface damage commonly causes plant leaves to release so-called green leaf volatiles, whereas blends of inducible compounds are more specific for herbivore attack and can vary considerably among plant genotypes. We compared the responses of naïve and experienced parasitoids of the species Cotesia marginiventris and Microplitis rufiventris to volatiles from maize leaves with fresh damage (mainly green leaf volatiles) vs. old damage (mainly terpenoids) in a six-arm olfactometer. These braconid wasps are both solitary endoparasitoids of lepidopteran larvae, but differ in geographical origin and host range. In choice experiments with odor blends from maize plants with fresh damage vs. blends from plants with old damage, inexperienced C. marginiventris showed a preference for the volatiles from freshly damaged leaves. No such preference was observed for inexperienced M. rufiventris. After an oviposition experience in hosts feeding on maize plants, C. marginiventris females were more attracted by a mixture of volatiles from fresh and old damage. Apparently, C. marginiventris has an innate preference for the odor of freshly damaged leaves, and this preference shifts in favor of a blend containing a mixture of green leaf volatiles plus terpenoids, after experiencing the latter blend in association with hosts. M. rufiventris responded poorly after experience and preferred fresh damage odors. Possibly, after associative learning, this species uses cues that are more directly related with the host presence, such as volatiles from host feces, which were not present in the odor sources offered in the olfactometer. The results demonstrate the complexity of the use of plant volatiles by parasitoids and show that different parasitoid species have evolved different strategies to exploit these signals.     

An elicitor in caterpillar oral secretions that induces corn seedlings to emit chemical signals attractive to parasitic wasps

Regurgitate of corn-fed beet armyworm (BAW) caterpillars,Spodoptera exigua, when applied to damaged sites of corn (Zea mays) seedlings, causes the release of relatively large amounts of terpenes by the seedlings several hours later. This plant response could be induced by merely placing the cut stem of seedlings in a solution of BAW regurgitate for 12 hr, a response that could not be induced by placing seedlings in water only. Regurgitate of BAW fed various diets, including a minimal diet of filter paper, were all active. However, seedlings placed in corn leaf juice, BAW hemolymph, or BAW feces extract released significantly smaller amounts of terpenes than did seedlings placed in BAW regurgitate. These results indicate that the active components are present in relatively large concentrations in regurgitate and that they are not related to the food source. Furthermore, regurgitate from several other species of caterpillars (Spodoptera frugiperda, Helicoverpa zea,Trichoplusia ni, andAnticarsia gemmatalis) as well as from the grasshopperSchistocerca americana induced the release of significant amounts of terpenes in corn seedlings. The release of these volatiles, therefore, appears to be a general response to attack by phytophagous insects. The terpene-releasing corn seedlings were highly attractive to the generalist parasitoidCotesia marginiventris and to the specialized parasitoidMicroplitis croceipes. This study confirms a systemic herbivore-elicited release of terpenes in corn. It is proposed that such chemicals serve multifunctional purposes that directly and indirectly protect plants against herbivorous arthropods and pathogens.     

β-Phellandrene: kairomone for pine engraver,Ips pini (Say) (Coleoptera: Scolytidae)

The responses ofIps pini (Scolytidae) to multiple-funnel traps baited with the pheromone, ipsdienol, and various monoterpenes were determined in stands of lodgepole pine in southern and central British Columbia.Ips pini was attracted to both ipsdienol and -phellandrene, demonstrating that -phellandrene is a kairomone for this species.Lasconotus complex (Colydiidae) and aCorticeus sp. (Tenebrionidae) were attracted to both ipsdienol and -phellandrene. TheCorticeus sp. exhibited a synergistic response to the combination of ipsdienol and -phellandrene; the responses of the other two species to the combination were additive. The predators,Thanasimus undatulus, Enoclerus sphegeus, andE. lecontei (Cleridae), were attracted to ipsdienol-baited traps, whileMonochamus clamator (Cerambycidae) andDendroctonus ponderosae (Scolytidae) were attracted to -phellandrene. Attraction of all eight species increased with increasing release rates of ipsdienol and/or -phellandrene.     

Predator-induced diel vertical migration inDaphnia: Enrichment and preliminary chemical characterization of a kairomone exuded by fish

Planktivorous fish release a cue of an unknown chemical nature into the water that induces diel vertical migration inDaphnia as predator avoidance response. We used a bioassay to obtain information about the chemical nature of the kairomone. We present a method to enrich the cue from holding water of fish by sorbent extraction. The kairomone exuded byLeucaspius delineatus can be characterized as a nonolefinic low-molecular-weight anion of intermediate lipophilicity. The presence of amino groups in the molecule can be excluded, whereas hydroxy groups are essential for activity. Separation by HPLC yielded only one active fraction. The kairomones released byCarassius carassius andRutilus rutilus showed the same chemical characteristics, suggesting that the kairomones from different species of fish, sensed byDaphia, are very similar, if not identical.     

(5Z,9Z)-3-alkyl-5-methylindolizidines fromSolenopsis (Diplorhoptrum) species

The alkaloidal venom components of two species of thief ants,Solenopsis (Diplorhoptrum) species AA andS. (Diplorhoptrum)conjurata have been found to contain (5Z,9Z)-3-hexyl-5-methylindolizidine and a mixture of (5Z,9Z)-3-ethyl-5-methylindolizidine andcis-2-methyl-6-nonyl-piperidine,trans-2-methyl-6-nonylpiperidine,cis-2-methyl-6-undecylpiperidine, and hexadecanoic acid.Monomorium pharaonis was similarly investigated and found to contain the indolizidine and pyrrolidines previously described (Ritter et al., 1977b). Both indolizidines were synthesized along with their stereoisomers and separated by preparative gas chromatography. Spectral studies revealed the stereochemistry to be 5Z,9Z in both cases. The stereochemistry of 2-butyl-5-pentylpyrrolidine inM. phaeronis has also been established. Biosynthetic relationships are discussed.     

Induction of volatile emissions in maize by different larval instars of Spodoptera littoralis

Maize plants under attack by caterpillars emit a specific blend of volatiles that is highly attractive to parasitic wasps. The release of these signals is induced by elicitors in the caterpillar regurgitant. Studies suggest that plants respond differently to different herbivore species and even to different herbivore stages, thus providing parasitoids and predators with specific signals. We tested if this is the case for different larval instars of the noctuid moth Spodoptera littoralis when they feed on maize plants. Cut maize plants were incubated in diluted regurgitant from second, third, or fifth instar caterpillars. There were no differences in total amount released after these treatments, but there were small differences in the release of the minor compounds phenethyl acetate and -humulene. Regurgitant of all three instars contained the elicitor volicitin. To test the effect of actual feeding by the larvae, potted plants were infested with caterpillars of one of the three instars, and volatiles were collected the following day. The intensity of the emissions was correlated with the number of larvae feeding on a plant, and with the amount of damage inflicted, but was independent of the instar that caused the damage. We also used artificial damage to mimic the manner of feeding of each instar to test the importance of physical aspects of damages for the odor emission. The emission was highly variable, but no differences were found among the different types of damage. In olfactometer tests, Microplitis rufiventris, a parasitoid that can only successfully parasitize second and early third instar S. littoralis, did not differentiate among the odors of maize plants attacked by different instar larvae. The odor analyses as well as the parasitoid's responses indicate that maize odors induced by S. littoralis provide parasitoids with poor information on the larval developmental stage. We discuss the results in the context of variability and lack of specificity in odorous plant signals.     

Comparative study of hermit crab responses to shell-related chemical cues

The responses of Australian hermit crabs to two types of shell-related odor cues were compared. Introduction of gastropod snail odor elicited an increase in grasping of shells in all four species tested (Clibanarius infraspinatus, C. taeniatus, C. virescens, andDiogenes avarus). However, the magnitude of response in the case ofC. virescens was significantly lower than the responses of individuals of the other species. Individuals ofC. infraspinatus responded to introduction of conspecific hemolymph with an increase in both locomotion and the frequency of grasping of shells.C. infraspinatus also responded to the hemolymph of the other three species, and there were no differences in the magnitude of the responses depending upon the source of the hemolymph. When individuals ofC. infraspinatus were exposed to snail odor and conspecific hemolymph at the same time, the responses were indistinguishable from those shown to hemolymph alone.     

Behavioral and biological responses ofCotesia marginiventris to kairomones of the fall armyworm,Spodoptera frugiperda

Potential kairomone sources of the fall armyworm (FAW),Spodoptera frugiperda (J.E. Smith), were bioassayed with females ofCotesia marginiventris (Cresson) in Petri dishes (10 cm diam). MatedC. marginiventris females, ranging in age from 1 to 3 days exhibited the most intense bioassay responses to potential sources of kairomone. Contacting a FAW-damaged corn leaf modified the pattern of movement inC. marginiventris from random to one exhibiting a significant increase in klinokinesis. No significant differences were present in kairomone responses of nonconditioned and conditioned parasitoids and parasitoid response to kairomones did not change throughout the photophase (0800–1800 hr). Removal of one, both, or the first eight antennal segments reduced or eliminated the response of the parasitoid to kairomones. Female parasitoids did not exhibit a preference for corn leaves damaged by a particular fall armyworm instar and parasitization rates were highest in larvae 48 hr old.Submitted in partial fulfillment of the requirements for Doctor of Philosophy, Department of Entomology and Nematology, University of Florida, Gainesville.